Cases Of Laryngeal Squamous Cell Carcinoma Research Articles

The basaloid variant of squamous cell carcinoma of the larynx

IntroductionBasaloid squamous cell carcinoma (BSCC) is a rare subtype of squamous cell carcinoma (SCC). Because of its rarity, both clinical and prognostic features of this variant are not well known. ObjectiveIn this study, we aimed to determine the frequency of BSCC and other SCC variants among all laryngeal SCC cases, and to determine clinical and prognostic features of BSCC variant. Study design: retrospective cohort study. Evidence level: Level 2b. Material and methodsRecords of the patients who had laryngeal SCC surgically treated at our institute between 2007 and 2013 were retrospectively reviewed. ResultsAmong 198 subjects who had laryngeal SCC surgically treated, the frequency of the variants of SCC other than classical variant was 10.1% (20/198). The most common SCC variant was BSCC (6.6%). Eleven (84.6%) patients with BSCC were at an advanced stage at the presentation (p>0.05). The 3-year overall survival and disease-free survival rates were 63% and 53% respectively. ConclusionBSCC variant may be more common than previously reported. Since almost the half of patients experiences disease recurrence in the early period, multimodal treatment strategies should be employed at initial treatment, and a close follow-up is strongly recommended for this aggressive SCC variant.

Notch1 overexpression associates with poor prognosis in human laryngeal squamous cell carcinoma.

This study aimed to investigate the expression of Notch1 in human laryngeal squamous cell carcinoma (LSCC) tissues and its relationship to clinicopathologic characteristics as well as their prognostic value in LSCC. Samples from 106 patients with LSCC were analyzed for Notch1 expression by immunohistochemical staining. The relationship between Notch1 expression and clinicopathologic parameters was subsequently analyzed. Univariate analysis and multivariate analysis of patient survival were examined using the Kaplan-Meier method and Cox proportional hazards model, respectively. We found that Notch1 had positive expression in 71 of 106 cases of LSCC (66.98%), which was obviously higher than laryngeal normal tissues (P < .01) and significantly correlated with the clinical stage, lymph node metastasis, and histological grade (all Ps < .05). Univariate analysis revealed that Notch1 expression tended to show an unfavorable influence on overall survival (OS) and disease-free survival (DFS) (both Ps < .01). Multivariate analysis demonstrated that Notch1 was an independent prognostic factor for patients with LSCC (P < .05). These results reveal that Notch1 expression is a potential prognostic factor for malignant progression, metastasis, and survival of LSCC patients. Furthermore, it has been demonstrated that high expression of Notch1 was associated with unfavorable OS and DFS in LSCC patients.

Expression and significance of angiopoietin-2 and cyclin D1 in laryngeal squamous cell carcinoma and the correlation with prognosis

The aim of this study was to study the expression of angiopoietin-2 (Ang-2) and the cell cycle protein D1 (cyclin D1) in laryngeal squamous cell carcinoma (SCC), and its clinicopathological meaning. The expression of Ang-2 and cyclin D1 was analyzed in 116 cases of laryngeal SCC, five cases of atypical hyperplasia and five cases of vocal polyp tissues using the immunohistochemical streptavidin-perosidase (S-P) method. Ang-2 and cyclin D1 protein expression was not present in 5 cases of atypical hyperplasia or 5 cases of vocal cord polyps. However, the proportions of positive staining in well, moderately and poorly differentiated laryngeal SCC were 40, 66.7 and 100%, respectively, for Ang-2 and 50, 66.7, 100%, respectively, for cyclin D1, and were statistically significant (P<0.05). The expression of Ang-2 was positively correlated with cyclin D1 in the laryngeal SCC (r=0.5042; P<0.05). This showed that the tumor grading and cyclin D1 were independent factors affecting laryngeal SCC patient survival by the Cox regression model of risk factors proportion analysis. Ang-2 is synergistic with cyclin D1 in the development of laryngeal SCC. Ang-2 is associated with the metastasis of lymph nodes. Detection of both Ang-2 and cyclin D1 may possess clinical significance in evaluating the prognosis and guiding the clinical treatment of SCC.

Establishment and characterization of a novel HPV-negative laryngeal squamous cell carcinoma cell line, FD-LSC-1, with missense and nonsense mutations of TP53 in the DNA-binding domain

Laryngeal squamous cell carcinoma (LSCC) is a common malignancy in China; however, publically available LSCC cell lines are few and not established from Chinese populations. Hence, novel and well-characterized LSCC cell lines of Chinese origin are urgently needed to provide researchers with a comprehensive database for LSCC research. From 40 cases of LSCC, we established a novel cell line that was maintained for more than 100 passages in vitro and was found to have typical epithelial morphology and ultrastructure. In-depth characterization analysis revealed polyploidy in DNA content; a doubling time of some 24h; high tumorigenicity in immunodeficient mice; higher invasive potential and more sensitive to radiation and cisplatin compared with HeLa cell line; upregulated Ki67, Notch1, EGFR, and CK5 protein levels; negative infection of human papillomavirus (HPV) and mycoplasma; expression of head and neck squamous cell carcinoma (HNSCC) biomarkers; mutations of TP53 in exons 5 and 8; a near-triploid karyotype with complex structural aberrations; and dozens of dysregulated genes and miRNAs. Cell authentication testing by the American Type Culture Collection (ATCC) confirmed the human origin of this cell line. Our findings indicate that a novel and well-differentiated LSCC cell line recapitulating the primary tumor’s malignant characteristics is established and well characterized. It does not match any cell lines within the ATCC database and helps to elucidate the molecular pathogenesis of LSCC.

Laryngeal Squamous Cell Carcinoma in 4256 Women: A Population‐Based Analysis of Gender's Prognostic Implications

Objectives: 1) Describe a large series of female patients with laryngeal squamous cell carcinoma (SCC). 2) Determine if there are gender-related differences in clinicopathologic factors, outcomes, and prognostic indicators. Methods: All cases of laryngeal SCC diagnosed between the years 2000-2008 were extracted from the Surveillance Epidemiology and End Results database. Clinicopathologic data, disease-specific survival (DSS), and prognostic factors were compared between genders using chi-square, Kaplan-Meier, and multivariable regression analysis. Results: Of 21,666 patients, 4,256 (19.6%) were female. Supraglottic tumors were predominant in females (61%) (vs. 64% glottic tumors in males; P &lt; 0.001); and females with supraglottic SCC were less likely to present with locally-advanced disease or nodal metastases ( P &lt; 0.001). Overall, female gender was independently associated with improved DSS (hazard ratio [HR] 0.81, 95% confidence interval [CI] 0.74-0.89, P &lt; 0.001). Further analysis stratified by age subgroups revealed that the favorable prognostic implications associated with female gender were limited to patients diagnosed at age&lt;70y ( P &lt; 0.005). In this group, females carried a 27-32% lower risk of SCC-specific mortality on multivariable analysis ( P &lt; 0.005). Notably, the protective effective of female gender was lost after age at diagnosis exceeded 70y ( P &gt; 0.25). Prognostic factors were mostly similar within each gender subgroup; however, the negative prognostic implications of age were significantly greater in females. In fact, the risk of laryngeal SCC-specific death nearly tripled per annum in females (2.9% vs. 1.6% in males, P &lt; 0.01). Conclusions: There are significant gender-related clinicopathologic differences in laryngeal SCC. Females diagnosed at age&lt;70y have significantly better DSS than males; and mortality risk is highly attributable to age in women.

HPV in squamous cell carcinoma: An Immuno-histochemical study

Objectives: This study is performed to evaluate the immune-expression of human papillomavirus (HPV) in a series of 100 samples of squamous cell carcinoma (SCC) from different sites of the body and to correlate it with anatomical site and grade of the tumor, age and sex of the patients, grade of tumor and to compare our results with those of other studies. Patients and methods: There were 100 patients with primary SCC from larynx, cervix, oral cavity and skin. Samples were obtained in a pro and retrospective fashion from teaching hospitals and private laboratories in Mosul. During a period of seven months from July 2009 through January 2010. Immunoexpression assay was performed using immunohistochemically (IHC). Results: The patients' age ranged between 15 and 90 years, with a mean of 51.28 year. There were 62 males and 38 females. Only nine of the hundred cases were positive for HPV, 3 out of 45 cases of laryngeal SCC were positive (6.67%) and 22 of oral cavity SCC cases were positive (27.27%). All of the 23 cervical and the 10 skin cases were negative for HPV. HPV expression was significantly correlated with the primary anatomical site but not with the age and sex of the patients in laryngeal and oral cavity SCC. Conclusions: 1.) HPV was expressed in nine out of 100 cases of SCC, adopting immunohistochemistry (IHC) for its detection, six of the cases were in the oral cavity and three were in the larynx with none from cervix or skin. 2.) In the positive cases the only significant correlations were with the anatomical site and in relation to grade in the larynx. 3.) Our results are low when compared to more sensitive methods for HPV detection like Polymerase Chain Reaction (PCR), Southern Blot Hybridization (SBH) and In Situ Hybridization (ISH); however this may also be attributed to the relatively small number of cases and to the possibility that our cases of SCC are not HPV related.

Expression of HMGA1 and Ezrin in laryngeal squamous cell carcinoma

Conclusion: The overexpression of HMGA1 or Ezrin may contribute to the carcinogenesis, development, and metastasis of laryngeal squamous cell carcinoma (LSCC). Objective: To investigate the expression of HMGA1 and Ezrin in LSCC and analyze their clinical significance. Methods: The expression of HMGA1 and Ezrin was analyzed by immunohistochemistry (IHC) in 50 cases of LSCC. Thirty cases of laryngeal polyp and 30 cases of atypical hyperplasia of larynx were studied as controls. The expression of HMGA1 and Ezrin was analyzed by real-time PCR and by Western blot in 30 cases of LSCC; samples from adjacent normal epithelial tissues in 30 cases were studied as controls. Results: (1) IHC revealed that the positive rate of HMGA1 protein was 68.0% (34/50), 53.3% (16/30), and 13. 3% (4/30) in LSCC, atypical hyperplasia of larynx, and laryngeal polyp (p < 0.05), and the positive rate of Ezrin protein was 64.0% (32/50), 50.0% (15/30), and 23.3% (7/30) (p < 0.01), respectively. (2) Real-time PCR demonstrated that the mean relative mRNA expression levels of HMGA1 in LSCC and in normal tissues were 2.41 ± 0.40 and 1.05 ± 0.18, respectively (p < 0.01). The mRNA levels of Ezrin in LSCC and in normal tissues were 1.79 ± 0.27 and 1.04 ± 0.22, respectively (p < 0.05). (3) Western blotting revealed that the mean relative protein expression levels of HMGA1 in LSCC and in normal tissues were 1.73 ± 0.60 and 0.35 ± 0.17, respectively (p < 0.01). The protein levels of Ezrin in LSCC and in normal tissues were 1.82 ± 0.77 and 0.42 ± 0.20, respectively (p < 0.01).

Expression of astrocyte elevated gene-1 protein and its clinical significance in laryngeal squamous cell carcinoma

To assess the protein expression of astrocyte elevated gene-1 (AEG-1) in tissue specimens of laryngeal squamous cell carcinoma (LSCC), and to correlate its expression with clinicopathological parameters and prognosis in patients with LSCC. RT-PCR was used to assay the expression of AEG-1 mRNA in 13 pairs of LSCC tissues and their corresponding noncarcinoma epithelia. Immunohistochemistry was performed on paraffin-embedded tissue specimens to investigate the protein expression of AEG-1 in 88 cases of LSCC specimens and 15 cases of adjacent epithelial samples. The expression of AEG-1 mRNA was significantly increased in LSCC tissues compared to adjacent noncarcinoma epithelial tissues (0.81 ± 0.17 vs. 0.23 ± 0.10;t = 10.337, P < 0.001). Meantime, the positive rate of AEG-1 protein in 88 cases of LSCC was 87.5% (77/88). However, 15 cases of adjacent noncarcinoma epithelial merely demonstrated negative or mild expression of AEG-1 protein. AEG-1 overexpression was closely correlated with T stage (χ(2) = 6.289, P = 0.018), clinical stage (χ(2) = 11.049, P < 0.01), metastasis (χ(2) = 20.859, P < 0.01) and recurrence(χ(2) = 13.459, P < 0.01). The overall survival rates of patients with AEG-1 overexpression and low expression were 35.9% and 86.4%, respectively (χ(2) = 23.409, P < 0.01). Multivariate Cox regression analysis revealed that AEG-1 expression was an independent prognostic factor (P = 0.016). AEG-1 protein may play a critical role in the initiation and progression of LSCC, implicating its predictive value in prognosis.

Expressions of key molecules affiliated cytoskeleton in laryngeal squamous cell carcinoma and their implications for prognosis

To study the expressions of key assemblies of cytoskeleton, Fascin-1, Ezrin and Paxillin, in laryngeal squamous cell carcinoma (LSCC) and their correlation with clinicopathologic characteristics, cancer recurrence and survival of patients with LSCC. The expressions of Fascin-1, Ezrin and Paxillin proteins were detected by immunohistochemistry in 199 cases of LSCC. Unconditional Logistic regression model or Cox proportional hazards model was used for the analyses of recurrent risks and prognostic factors. Significantly increased expression of Fascin-1, Ezrin or Paxillin expression was showed in the LSCC with poorly differentiated, positively cervical lymph nodal metastasis, and clinical stage III + IV respectively (P < 0.05). The expressions of three kinds of proteins in the recurrent cases were higher than those in non-recurrent cases respectively (χ(2) were 42.479, 43.673 and 22.261, P < 0.05). The highest recurrence rate (69.1%) was observed in group of cases with the highly co-expression of the three kinds of proteins (P < 0.05). The expression of Fascin-1 (OR = 7.89, 95%CI 2.26 - 27.53, P = 0.001), or Ezrin (OR = 2.51, 95%CI 1.18 - 5.32, P < 0.001) was independent risk for recurrence. Five-year disease-free survival rates of patients with high expression of Fascin-1, Ezrin or Paxillin were lower than those of patients with negative or low expressions for the proteins (P < 0.05). Patients with highly co-expression of three kinds of proteins showed the poorest survival prognosis, with a 5-year disease free survival (DFS) of only 26.4% (P < 0.05), and expressions of three proteins were independent prognostic factors for 5-year DFS (P < 0.05). Fascin-1, Ezrin, and Paxillin were correlative with LSCC progression and might be potential predictors for cancer recurrence and survival of patients with LSCC, as well as therapeutic targets for LSCC.

Vasculogenic mimicry is a key prognostic factor for laryngeal squamous cell carcinoma: a new pattern of blood supply.

Recurrence and local lymph node metastasis affected the prognosis of patients with laryngeal cancer. The aim of this study was to analyze clinical and pathological significance of vasculogenic mimicry (VM) in laryngeal squamous cell carcinoma (LSCC) and evaluate its contribution to prognosis. Data of 168 cases of LSCC were reviewed retrospectively to reveal clinical pathology and prognostic significance of VM. CD31 and periodic acid-Schiff double staining was used to identify VM. VM in LSCC contributed to lymph node metastasis (P = 0.003) and clinical progression. VM correlated to histopathology grade (P = 0.001) of LSCC. VM was an adverse prognostic factor for both disease-specific survival (P = 0.039) and metastasis-free survival (P = 0.042) by univariate survival analyses. And it was an independent prognostic factor for only disease-specific survival (P = 0.003) by multivariate survival analyses. VM existed in LSCC. LSCC with VM has more potential to invasion and metastasis.

Expression of HMGB1 protein in laryngeal squamous cell carcinoma and its clinical significance

To evaluate the expression of HMGB1 protein in tissue specimens of laryngeal squamous cell carcinoma (LSCC) and adjacent normal mucosa, and explore the correlation of HMGB1 protein expression with clinicopathologic features and prognosis in LSCC. Ninty-three cases of LSCC and 5 cases of adjcent mucosal tissue samples were included in this study. Immunohistochemical staining was performed on paraffin-embedded tissue specimens to examine the HMGB1 protein expression. The data were futher correlated with the clinicopathological features and prognosis of the LSCC patients. The positive rates of HMGB1 expression in LSCC specimens was 87.1%, significantly higher than that in the adjcent normal mucosa samples (46.7%, P = 0.001), and its overexpresion was closely correlated with T stage (Chi2 = 10.878, P = 0.004), clinical stage (Chi2 = 21.115, P < 0.01), metastasis (Chi2 = 28.298, P < 0.01) and recurrence (Chi2 = 14. 923, P = 0.001) in patients with LSCC. Patients with HMGB1 overexpression had both poorer disease-free survival and poorer overall survival compared with that in patients with low HMGB1 expression (Chi2 = 13.815, Chi2 = 11.912; Both P < 0.01). Univariate and multivariate Cox regression analyses revealed that HMGBI expression is an independent prognostic factor for patients with LSCC. The results of this study demonstrate that HMGB1 protein expression is significantly increased in LSCC tissues, and HMGB1 protein overexpression is associated with a poorer prognosis in patients with LSCC. These results suggest that HMGB1 may play a critical role in the initiation and progression of LSCC, implicating HMGB1 may become a valuable marker for the prediction of prognosis in patients with LSCC.

Analysis of Transcriptional Factors and Regulation Networks in Laryngeal Squamous Cell Carcinoma Patients with Lymph Node Metastasis

The present study was to identify and quantitate differentially expressed proteins in laryngeal squamous cell carcinoma (LSCC) tissues with or without lymph node metastasis and to explore transcriptional factors and regulation networks associated with the process. Tissue specimens were taken from 20 patients with LSCC, including 10 cases of LSCC without metastasis LSCC (N0) and 10 cases of LSCC with metastasis LSCC (Nx). Among the 643 unique proteins identified by using iTRAQ labeling and quantitative proteomic technology, 389 proteins showed an abundance change in LSCC (Nx) as compared to LSCC (N0). Cytoskeleton remodeling, cell adhesion, and immune response activation were found to be the main processes in LSCC metastasis. The construction of transcription regulation networks identified key transcription regulators for lymph node metastasis of LSCC, including Sp1, c-myc, and p53, which may affect LSCC metastasis through the epithelial-mesenchymal transition. Furthermore, our results suggest that ubiquitination may be a critical factor in the networks. The present study provides insights into transcriptional factors and regulation networks involved in LSCC metastasis, which may lead to new strategies for treatment of LSCC metastasis.

Prognostic significance of matrix metalloproteinase-20 overexpression in laryngeal squamous cell carcinoma

Conclusion: Matrix metalloproteinase (MMP)-20 is overexpressed in laryngeal squamous cell carcinoma (LSCC) compared with the adjacent normal laryngeal epithelium and MMP-20 plays a role in lymph node metastasis. Overexpression of MMP-20 may be used as a significant prognostic factor for lymph node metastasis. All the findings indicate that MMP-20 may play a role in the initiation and progression of LSCC. Objective: The MMPs are a gene family of zinc-dependent endopeptidases that have been implicated in tumor invasion and metastasis, and MMP-20 is a new member of the MMP family. The purpose of this study was to investigate whether MMP-20 is overexpressed in human LSCC and, if so, the significance of its overexpression in relation to clinical parameters. Methods: We analyzed 33 cases of LSCC with RT-PCR and 73 cases of LSCC with immunohistochemistry compared with normal laryngeal epithelium. Results: We found that MMP-20 is overexpresssed in LSCC compared with the adjacent normal laryngeal epithelium.

Vasculogenic mimicry contributes to lymph node metastasis of laryngeal squamous cell carcinoma

BackgroundSurvival of laryngeal squamous cell carcinoma (LSCC) patients has remained unchanged over recent years due to its uncontrolled recurrence and local lymph node metastasis. Vasculogenic mimicry (VM) is an alternative type of blood supplement related to more aggressive tumor biology and increased tumor-related mortality. This study aimed to investigate the unique role of VM in the progression of LSCC.MethodsWe reviewed clinical pathological data of 203 cases of LSCC both prospectively and retrospectively. VM and endothelium-dependent vessel (EDV) were detected by immunohistochemistry and double staining to compare their different clinical pathological significance in LSCC. Survival analyses were performed to assess their prognostic significance as well.ResultsBoth VM and EDV existed in LSCC type of blood supply. VM is related to pTNM stage, lymph node metastasis and pathology grade. In contrust, EDV related to location, pTNM stage, T stage and distant metastasis. Univariate analysis showed VM, pTNM stage, T classification, nodal status, histopathological grade, tumor size, and radiotherapy to be related to overall survival (OS). While, VM, location, tumor size and radiotherapy were found to relate to disease free survival (DFS). Multivariate analysis indicated that VM, but not EDV, was an adverse predictor for both OS and DFS.ConclusionsVM existed in LSCC. It contributed to the progression of LSCC by promoting lymph node metastasis. It is an independent predictors of a poor prognosis of LSCC.

Laryngeal carcinoma lymph node metastasis and disease-free survival correlate with MASPIN nuclear expression but not with EGFR expression: a series of 108 cases

Despite advances in laryngeal squamous cell carcinoma (LSCC) treatment, patient survival has not improved in the last two decades. Novel, more effective strategies should be based on receptor-mediated LSCC-targeted therapy. The epidermal growth factor receptor (EGFR) is the most widely studied molecular target. MASPIN multifaceted anti-tumor effects have been rarely evaluated in LSCC. The aims of this study were to investigate EGFR and MASPIN expression and the role of sub-cellular MASPIN localization in LSSC. MASPIN and EGFR expression and the sub-cellular localization of MASPIN were assessed using a computerized image analysis system in 108 consecutive cases of operable LSCC. The rates of occurrence of lymph node metastases and recurrent disease were lower in patients with a nuclear pattern of MASPIN expression (p = 0.004, p = 0.0028). As expected, the loco-regional recurrence rate was lower in N0 patients (p = 0.009), but the disease recurrence rate was even lower in N0 patients with a nuclear localization of MASPIN (p = 0.020). Disease-free survival was longer in cases of LSCC with a nuclear MASPIN pattern (p = 0.003). The intensity of EGFR reactivity and the EGFR area fraction were unrelated to the clinico-pathological and prognostic parameters in LSCC. A nuclear MASPIN pattern is a promising prognostic indicator in LSCC, but further evidence is needed before elective neck dissection can be considered for cN0 LSCCs with a non-nuclear MASPIN pattern. Although a better understanding of the mechanisms behind sub-cellular MASPIN localization is mandatory, re-activating nuclear MASPIN in association with specific chemotherapeutic agents may be an important goal in the treatment of LSCC.

Expression of human beta-defensin-2 in laryngeal squamous cell carcinoma and its correlation with CD1a(+) dendritic cells infiltrating in tumor.

To investigate the expression of human beta-defensin-2 (HBD-2) in laryngeal squamous cell carcinoma (LSCC), and reveal the immune significance of HBD-2 in LSCC. The expression of HBD-2 and the infiltration of CD1a(+) dendritic cells were verified by immunohistochemistry strept actividin-biotin complex (SABC) method in thirty-four cases of LSCC's paraffin sections, and the expression of HBD-2 mRNA was detected in eighteen cases of postoperative tumor specimens and five cases of normal paracarcinoma tissue using cDNA probe by in situ hybridization. Based on the type and characteristic of datas, SPSS13.0 program package was used for statistical calculation. (1) The difference of expression of HBD-2 was significant among the grades of malignant cell differentiation (F = 6.809, P < 0.05), the expression of HBD-2 in well differentiated group was significantly increased compared with moderately and poorly differentiated group (P < 0.05), there wasn't significant difference with the clinical type, T status and lymphatic metastasis in the expression of HBD-2. (2) The difference of expression of HBD-2 mRNA among different grades of malignant cell differentiation was significant (F = 16.391, P < 0.05), less or no positive signal was observed in poorly differentiated group. (3) The infiltration of CD1a(+) dendritic cells had positive correlation with the expression of HBD-2 (r = 0.343, P < 0.05). The expression of HBD-2 was related to the grade of malignant cell differentiation in LSCC, which was regulated at the level of transcription and translation. HBD-2 might play a role in anti-tumor immunity by chemo-attractiving and activating of immature dendritic cells.

Suppression of Survivin Expression by Short Hairpin RNA Induces Apoptosis in Human Laryngeal Carcinoma Cells

Purpose: The aim of this study was to evaluate the suppression effect of survivin shRNA on the expression of the survivin gene in the human laryngeal cancer cell line Hep-2. Procedures: 60 cases of laryngeal squamous-cell carcinoma (LSCC) and 10 cases of normal laryngeal mucosa were examined using immunohistochemistry to determine whether the expression of survivin correlated with tumorigenesis. Three plasmid vectors of short hairpin RNA (shRNA) specific for survivin were designed and generated. Western blot and real-time PCR analysis of survivin expression in Hep-2 cells was performed 48 h after transfection. The growth curve was used to determine the cell proliferation. Propidium iodide (PI) single staining was applied to detect the cell cycle. The apoptosis of the cells was analyzed by flow cytometry with the FITC-annexin-V/PI double staining and PI single staining. Results: 68.33% (41 out of 60) of tumors were positive for survivin expression and significantly associated with lymph node metastasis and advanced stage. In contrast, no expression of survivin in normal mucosa was detected. Transfection of Hep-2 cells with survivin shRNA significantly inhibited survivin expression at both the mRNA and the protein level in Hep-2 cells. Downregulation of survivin resulted in increasing the apoptosis index, but the results showed no obvious influence on cell cycle. Conclusions: This study demonstrates that survivin shRNA effectively inhibits survivin gene expression in Hep-2 cells leading to growth suppression and apoptotic induction in Hep-2 cells.

Expression of cyclin G1 in laryngeal squamous cell carcinoma and its clinicopathologic significance

To evaluate the expression of cyclin G1 and its relationship with clinical pathological characters, p53 expression and microvessel density (MVD) in laryngeal squamous cell carcinoma (LSCC), and to investigate the prognostic value of cyclin G1 in LSCC. Using immunohistochemistry, 81 cases of LSCC were evaluated for cyclin G1 and p52 expression, MVD was assessed by CD34 staining. In 20 cases, similar assessment was made in the adjacent normal mucosa. Compared to the normal laryngeal mucosa, the expressions of cyclin Gl, p53 and MVD were higher in LSCC than that in normal tissues. The positive rates of cyclin G1 and p53 expressions in LSCC were 61.7% (50/81) and 65.4% (53/81), respectively. The positive rates of cyclin G1 in LSCC with or without p53 expression were 71.7% (38/53) and 42.9% (12/28), respectively. The expression of cyclin G1 was significantly correlated with p53 (P = 0.011). The mean value of MVD was (51.23 +/- 16.46 mv) in cyclin G1 positive group, and(30.74 +/- 12.29 mv) in the negative group, and the difference was significant (P = 0.005). Furthermore, the over-expression of cyclin G1 was related to the histological grade, cervical lymph node metastases and 5 year survival (P = 0.002, 0.013 and 0.032 respectively). Cyclin G1 is highly expressed in laryngeal squamous cell carcinoma, and is associated with the expression of p53 protein and tumor angiogenesis. It may play an important role in carcinogenesis and metastasis in LSCC, and may be one of prognostic indices for LSCC patients.

Expression of cyclooxygenase-2, vascular endothelial growth factor and matrix metalloproteinase-2 in patients with primary laryngeal carcinoma: a tissue microarray study

To determine the correlation between expression of cyclooxygenase-2, vascular endothelial growth factor and matrix metalloproteinase-2, in patients with laryngeal carcinoma. The study included 85 primary laryngeal squamous cell carcinoma cases. Expression was assessed using Envision immunohistochemical stains for cyclooxygenase-2, vascular endothelial growth factor and matrix metalloproteinase-2. A tissue microarray containing samples from the 85 primary laryngeal squamous cell carcinoma cases was assembled. Immunohistochemical testing for cyclooxygenase-2, vascular endothelial growth factor and matrix metalloproteinase-2 was performed. Using Pearson correlation, expression of these proteins was compared with the following clinicopathological variables: age, sex, clinical tumour-node-metastasis staging, and prognosis. Three-year survival curves, factored by cyclooxygenase-2, vascular endothelial growth factor and matrix metalloproteinase-2 expression, were generated for overall survival, by Kaplan-Meier analysis. The expression of cyclooxygenase-2 significantly differed between patients with different pathology, tumour-node-metastasis stage and prognosis. A marked difference in vascular endothelial growth factor expression was seen between two histological grade groups. Expression of matrix metalloproteinase-2 protein statistically significantly differed between patients with different tumour-node-metastasis stages, lymph node metastases and three-year survival rates. The expression of cyclooxygenase-2 in laryngeal carcinoma tissue was found to be associated with the expression of matrix metalloproteinase-2. Cyclooxygenase-2 and matrix metalloproteinase-2 may act as clinical prognostic indicators of tumour growth and differentiation in laryngeal carcinoma.

Human papilloma virus prevalence in laryngeal squamous cell carcinoma

To determine the prevalence and type of human papilloma virus deoxyribonucleic acid (DNA) in cases of laryngeal squamous cell carcinoma. We analysed the prevalence of human papilloma virus infection in archived paraffin block specimens taken from 99 cases of laryngeal squamous cell carcinoma between 1990 and 2005, using polymerase chain reaction techniques. Biopsy specimens from five proven verrucous skin lesions were used as positive controls, and peripheral blood samples from five healthy volunteers were used as negative controls. Four test samples were found to have inadequate deoxyribonucleic acid purity and were therefore excluded from the study. Human papilloma virus deoxyribonucleic acid was detected in seven of 95 cases of laryngeal squamous cell carcinoma (7.36 per cent). Human papilloma virus genotyping revealed double human papilloma virus infection in three cases and single human papilloma virus infection in the remaining four cases. The human papilloma virus genotypes detected were 6, 11 and 16 (the latter detected in only one case). In our series, a very low human papilloma virus prevalence was found among laryngeal squamous cell carcinoma cases. The human papilloma virus genotypes detected were mostly 6 and/or 11, and 16 in only one case. To the best of our knowledge, this is the first report of human papilloma virus prevalence in laryngeal squamous cell carcinoma, based on polymerase chain reaction genotyping in a Turkish population.